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I.J.S.N., VOL. 1(1):38-41
ISSN 0973 -3140
EFFECTS OF CRUDE EXTRACTS OF ABELMOSCHUS ESCULENTUS ON
ALBUMIN AND TOTAL BILIRUBIN OF DIABETIC ALBINO RATS
Uraku, A. J1., Ajah, P. M1., Okaka, A. N. C2, Ibiam, U. A1 and *Onu, P. N3
1
Department of Biochemistry/Biotechnology, Ebonyi State University.
Department of Biochemisrty Nnamdi Azikiwe University Awka, Anambra State, Nigeria.
3
Department of Animal Science, Ebonyi State University, PMB 053 Abakaliki, Ebonyi State Nigeria.
*Corresponding author email: [email protected]
2
ABSTRACT
The effects of crude extracts of A. esculentus on albumin and total bilirubin levels of diabetic albino rats were investigated.
Sixty (60) albino rats were divided into four (4) groups of namely A, B, C and D. While A and B groups served as positive
and negative controls, respectively, groups C and D served as the experimental groups and each was further subdivided
into three (3) groups of 5 animals each. Hyperglycemia was induced in the treatment sub-groups by a single intraperitoneal
injection of alloxan at 75 mg per kg body weight. The extracts were administered via oral intubation to the subgroups of C
and D in and doses of 200,400 and 800 mg/kg body weight respectively for a period of 14 days. Blood samples were
collected through tail vein on various stages and serum glucose, albumin and total bilirubin concentrations were assayed
using spectrophotometric method. Results showed that administration of A. esculentus extract to diabetic rats group
recorded significantly higher (p<0.05) body weight gain than those of the test-control and control. There was a significant
(p<0.05) increase in serum glucose of the untreated diabetic group compare to the control group. Administration of
aqueous-ethanolic extracts caused a marked (p<0.05) increased in the albumin levels in the diabetic rats. There was a
significant (P<0.05) increase (82%) in total bilirubin levels in diabetic control group over the normal control.
Administration of aqueous-ethanolic extracts caused a marked significant (p<0.05) decrease in the billrubin levels in the
diabetic rats.
KEY WORDS: Abelmoschus esculentus, Diabetic mellitus, hypoglycemic effect, Serum albumin and total bilirubin.
INTRODUCTION
Plant has been the source of help, survival and good
health. Plants are able to be functioning in this capacity
because they have many important chemical substances
found all over their various parts such as alkaloids, carbon
compounds, nitrogen, glycosides, essential oils, fatty oils,
resins, mucilage, tannins, gums and others (Osmund,
2001). Most of these are potent bioactive compounds
found in medicinal plant parts that can be used for
therapeutic purpose or which are precursors for the
synthesis of useful drugs (Sofowora, 1993).
The active principles differ from Plants to plants due to
their biodiversity and they produce a definite physiological
action on the human body. Also, there is growing interest
on the medicinal properties of a number of common plants
(Gbile and Adosina, 2003). Lawal et al. (2006) have
elucidated the importance of these medicinal plants and
their importance in the pharmaceutical industries. These
plants have been under utilized in orthodox medicine but
have continued to be used in ethno-medicinal preparations.
Today about 300 species of medicinal and aromatic plants
are used worldwide in the pharmaceuticals, foods,
cosmetics and perfume industries (Osmund, 2001; kafaru,
1984). Abelmoschus. esculentus, a flowering plant in the
mallow family mavaceae, originating somewhere near
present day Ethiopia (Abyssina) (Keay, 1989) but spread
to the other continents via African slave trade route (Keay,
1989: Schipper,2000). It was formally considered a
species of Hibiscus but now classified in the genus of
Abelmoschus. It was grown throughout the tropical and
warm temperate regions of the world (Schipper, 2000).
Due to the nutrient content of the A. esculentus, the pods,
flowers, leaves and fruits are used as therapeutic diets. In
countries such as Turkey and Cyrus, the plants are used in
preparing medicinal remedies to and reduce swelling;
inflammation (Olusola et al., 2006). The consumption has
been reported to reduce serum cholesterol, triacylglyceride
and blood pressure (Reid, 1990: Andullu and
Varderchyulu, 2001). In some part of Africa such as
Ethiopia, the parts are used as part of therapeutic diet
against menstrual pains and for hypertension (Daly, 1997).
In the preset study we investigated the effects of the
ethanoic and aqueous extracts of A. esculentus fruits on
serum glucose, albumin and total bilirubin in albino rats.
MATERIALS AND METHODS
Collection and Preparation of A.esculentus Extracts
Young fruits of A. esculentus were collected from Ogboji Agoutu in Inyaba Development Centre of Ebonyi State,
Nigeria and identified by a plant taxonomist in the
Department of Zoology, Ebonyi State University,
38
Effects of Abelmoschus esculentus on albumin and total bilirubin on diabetic rats
Abakaliki. They were washed thoroughly under running
tap water, shade dried and pulverized, using a grinding
machine. Aqueous and ethanolic extracts were prepared by
soaking about 100g of the powdered leaves in 1500ml and
800ml of water and ethanol, respectively for 24hours. The
extract was filtered and the solvents removed with rotary
evaporations to obtain crude active ingredient.
Group D2 : Diabetic rats given ethanolic extract of A.
esculentus (400mg/kg body weight) daily using a canular
for 14 days.
Group D3: Diabetic rats given ethanolic extract of A.
esculentus (800mg/kg body weight) daily using a canular
for 14 day Blood samples were collection from the rats at
various stages of the experiment namely, at the initial
stage after acclimatization, 72hours after injection of
alloxan and 14 days after administration of the extracts.
Blood samples were collected from the animal via the tail
vein under mild anesthesia with chloroform.
Experimental Animal
Sixty (60) albino rats of both sexes each weighing 122.50
– 165.50g were procured from The Pharmacy Department
of University of Nigeria, Nsukka, Nigeria. The animals
were acclimatized for 7 day under standard environment
conditions and fed ad-libitum on their normal diets. All
animals were fasted before the start of the experiment. The
animals were distributed into four (4) groups (A, B, C and
D). Groups C and D were subdivided into three (3) groups
with each sub-group having four animals. Group A was
Normal control, Group B, Diabetic control, Group C:
Diabetic rats treated with aqueous extract of
A.
esculentus and Group D: Diabetic rats treated with
ethanolic extract of A. esculentus.
Each animal for diabetic assay was induced by a single
intraperitoneal injection of alloxan solution at a dose of
75mg/kg body weight after an overnight fast to groups B,
C and D. However, the animals in group A did not receive
alloxan dosage and served as animal control. The blood
glucose level of the animals was checked using a
glucometer (a one touch test strips) after alloxan injection
using a method described by Tietz (2000). The blood
glucose level of the animals were again checked after 7
days to ascertain a diabetic state, and rats with moderate
diabetes were used for the experiment.
Determination of Serum Glucose, total Bilirubin and
Albumin
Serum glucose levels were determined in rats using a
method described by Tietz (2000).Total serum bilirubin
was determined using a modification Tietz (2000) and
Jendrassik and Grof (1938). Spectroscopic determination
of serum albumin levels was carried out based on a
modification of Tietz (2000) and Grant et al., (1987).
Statistical Analysis
All the results obtained were expressed as mean ± S.D of 5
rats in each group.All the tested parameters were subjected
to statistical analysis using ANOVA. Differences between
means were regarded significant at P<0.O5
RESULTS
The effect of the aqueous and ethanolic extracts of A.
esculentus on the Weight changes and serum glucose
levels of albino rats is shown in Table 1. There was a
significant (p<0.05) decrease in weight gain of the
untreated diabetic group compare to the control group.
Administration of A. esculentus extract to diabetic rats
group recorded significantly higher (p<0.05) body weight
gain than those of the test-control and control. There was a
significant (p<0.05) increase in serum glucose of the
untreated diabetic group compare to the control group.
Administration of aqueous-ethanolic extracts Abelmoschus
esculentus at 200, 400 and 800 mg/kg to the alloxan
induced diabetic rats caused a significant (p<0.05)
decrease in fasting serum glucose levels of the rats.
Administration of aqueous extract caused a marginal
(p<0.05) reduction in serum glucose of the treated diabetic
group compare to the ethanolic extracts treated group
Effect of the aqueous and ethanolic extracts of A.
esculentus on the serum albumin and total bilirubin levels
of albino rats is shown in Table 2. The treated animals had
significantly lower (p<0.05) plasma albumin levels,
compared to the normal-control. Albumin level also
decreased significantly (p<0.05) by 43% in diabetic
control group compared to normal control group.
Administration of aqueous-ethanolic extracts caused a
marked (p<0.05) increased in the albumin levels in the
diabetic rats. There was a significant (P<0.05) increase
(82%) in total bilirubin levels in diabetic control group
over the normal control. Administration of aqueousethanolic extracts caused a marked significant (p<0.05)
decrease in the billrubin levels in the diabetic rats.
Experimental design: In this experiment, a total of 56
albino rats (20 diabetic surviving rats, 10 normal rats)
were used. Diabetes was induced in rats a week before the
start of the experiment. The rats were divided into four
groups after the induction of diabetes. Varying
concentrations of the crude extracts of A. esculentus were
administered via oral intubation to the animals in groups C
and D subgroups ( C1, C2, C3 and D1, D2, and D3) for a
period of 14 days. These served as diabetic experimental
groups while those in group B did not receive the extracts
and served as diabetic control.
Group A : Normal untreated rats.
Group B : Diabetic untreated rats
Group C1 : Diabetic rats given aqueous extract of A.
esculentus (200mg/kg body weight) daily using a canular
for 14 days
Group C2 : Diabetic rats given aqueous extract of A.
esculentus (400mg/kg body weight) daily using a canular
for 14 days.
Group C3 : Diabetic rats given aqueous extract of A.
esculentus (800mg/kg body weight) daily using a canular
for 14 day
Group D1 : Diabetic rats given ethanolic extract of A.
esculentus (200mg/kg body weight) daily using a canular
for 14 days
39
I.J.S.N., VOL. 1(1):38-41
ISSN 0973 -3140
Table 1: Weight changes and serum glucose levels of albino rats after administration of A. esculentus extracts
Groups
Initial weight (g)
Final weight (g)
Initial glucose(mg/dl)
Final
glucose(mg/dl)
A
165.50 ± 26.73
168.0 0 ±26.50
89.50 ± 2.50
94.00 ± 3.67
B
122.50 ± 4.94
93.75 ± 4.15*
223.75 ± 16.35
241.25 ± 17.46
C1
165.50 ± 16.45
193.75 ± 14.31#
231.25 ± 7.40
208.75 ± 9.60#
#
C2
155.00 ± 18.37
185.75 ± 15.63
232.50 ± 12.50
197.50 ± 11.46#
#
C3
148.75 ± 16.45
187.50 ± 4.82
233.70 ± 9.60
195.00 ± 12.75#
#
D1
139.50 ± 10.31
167.25 ± 13.92
232.50 ± 5.59
217.50 ± 10.54#
#
D2
154.25 ± 8.61
178.75 ± 2.17
228.75 ± 5.45
202.50 ± 5.59#
#
D3
160.75 ± 14.96
188.25 ± 13.20
235.00 ± 9.35
200.00 ± 13.69#
Values are mean ± SD, n = 5 animals per group. *: Significantly different from the control group (p<0.05). #:
Significantly different from the diabetic group (p<0.05).
Table2: Serum albumin and total bilirubin levels of albino rats after administration of A. esculentus.
Parameters
Groups
Albumin (g/dl)
Total bilirubin(mg/dl)
A
10.18 ± 0.55
1.98 ± 0.33
B
5.03 ± 0.48*
3.60 ± 0.32*
C1
6.15 ± 0.48#
2.93 ± 0.29#
#
C2
6.33 ± 0.44
2.88 ± 0.24#
#
C3
6.40 ± 0.44
2.70 ± 0.19#
#
D1
5.98 ± 0.47
3.03 ± 0.29#
#
D2
6.10 ± 0.51
2.90 ± 0.31#
#
D3
6.18 ± 0.52
2.87 ± 0.33#
Values are mean ± SD, n = 5 animals per group. *: Significantly different from the control group (p<0.05). #:
Significantly different from the diabetic group (p<0.05).
uncontrolled hyperglycemia in alloxan induced animals.
Administration of aqueous-ethanolic extracts Abelmoschus
esculentus at 200, 400 and 800 mg/kg to the alloxan
induced diabetic rats caused a significant (p<0.05)
decrease in fasting serum glucose levels with percentage
values of 13 and 10%; 18 and 16%; 19 and 17%,
respectively. This suggests that the extracts contain
principles with the potential of reducing glucose level in
serum. The marginal reduction in serum glucose achieved
with aqueous extract at the various levels indicates that
aqueous extract is more effective in reducing srum glucose
level than ethanolic extract. This observation is in
agreement with the report of Iwueke et al. (2006) that
aqueous extract was more efficient in reducing blood
glucose level than the ethanolic extract.
Albumin level also decreased significantly (p<0.05) by
43% in diabetic control group compared to normal control
group. This observation may be attributed to inducement
of hyperglycemia by alloxan. Hyperglycemia increased
gluconeogenesis and thereby resulting to excess protein
breakdown and also excess nitrogen loss due to
gluconeogenesis (Robert et al., 2000).
However, administration of aqueous-ethanolic extracts
caused a remarkable increased in the albumin levels in the
diabetic rats with percentage change values of 22 and
19%; 26 and 21%, and 27 and 30 % for 200, 400 and 800
mg/kg weight doses, respectively. These observations may
be due to the presence of some compounds which help in
provision of a reserved store of protein (Nsirim, 1999;
Kaplan et al., 1999). However, the extracts mechanisms
DISCUSSION
The effectiveness of ethanolic and aqueous extracts of the
leaf of Abelmoschus esculentus in reducing blood glucose
level in diabetic rats and its effects on blood albumin and
bilirubin levels were investigated and compared.The
percentage yields of the ethanolic and aqueous extracts
were 21.00 and 25.00 respectively. This shows that a
substantial fraction of the constituents is freely soluble in
ethanol and water. This could explain the use of the
aqueous extracts of most medicinal plants by traditional
medicine practitioners for the sole purpose of achieving
high yield and potency.
The observed significant decrease in the mean body
weight in group B may be ascribed to the reduction in
utilization of food and fluid. However, Induction of
hyperglycemia has been linked to weight loss (Nelson and
Cox, 2000). The animals in experimental groups recorded
significant (p< 0.05) weight gains compared to diabetic
control groups (Table 1). Plant extracts effects may be due
to a greater efficiency in the utilization of feed, resulting in
enhanced growth. There is evidence to suggest that plant
extracts have appetite- and digestion-stimulating
properties and antimicrobial effects. This observation
agrees with the report of Agbafor and Akubugwo,
2006which state that the increase in mean body weight of
animals given lemon grass is a result of increase in
utilization of food and fluid.
A significant (p< 0.05) elevation of 150% and 157% in
serum glucose was observed in diabetic control when
compared with normal control. This increase indicates
40
Effects of Abelmoschus esculentus on albumin and total bilirubin on diabetic rats
of action behind these observations remain to be
determined although the mechanism suggested here may
not be ruled out completely.
There was a 82% increase in total bilirubin levels in
diabetic control group. On treatment with the extracts, the
levels decreased by 19 and 16%; 20 and 26 and, 22 and
20% (P<0.05) of that of diabetic control. This indicates
that hyperglycemia can enhance protein glycation. The
increased total bilirubin levels indicate that excess
haemoglobin is being destroyed or that the liver is not
actively treating the haemoglobin it is receiving (Andullu
and Vardycheryalu, 2001 and Tietz, 2000).
In conclusion, the results of this study suggest that the
fruits of Abelmoschus esculentus
extracts possess
hypoglycemic potential. However, aqueous extracts is
more effective in controlling gluconeogenesis.
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